I am working with wood decay fungi. I want to make a homogenize mycelium suspension, I used waring blender for different times, But I have lumps of mycelium every time which don't want. I want to impregnate wood blocks with this mycelium suspension.
if any body have any suggestion, I will appreciate your answers.
HI Nasim
For making homogenized fungal culture suspension, first grow your fungi in broth culture in shaking condition (app. 150 rpm) and afterward centrifuge the culture for breaking down of mycelium and filter through cheese cloth.
Good Luck
Rather than using a standard blender attachment, use a mixing/grinding attachment or use a food processor. The mixing/grinding attachments have a different blade position than standard blenders which reduce large clumps. You might still need to filter through cheesecloth, but for many of our fungi we do not have to filter.
Dear Nasim,
Two questions:
What species do you study?
Did you try to find out the nature of the mycelial lumps by light microscopy? That could clarify if you need to filter your suspension or work with lumps
Hi Nasim,
The only way to really get a homogeneous mycelial suspension is the use of a homogenizer. It is true, however, that they are quite expensive.
Regards, Franci
Dear Maria,
i am working with wood decaying fungi like Gleophylum trebium and Irpex lacteus, i want to inoculate my wood wafers with the same amount of mycelium suspension because all the wood blocks should contaminate at the same rate. then I will examine the volatile that fungi will produce during the metabolites. at first I tried to collect the spores from my fungi but unfortunately this type of fungi are not producing spore in-vitro easily, now I want to use homogenized mycelium.
Hi Swati
You can remove the mycelium and make a shaker for the broth that contain mycelium and spores, then you can do a filtration for it.
Hi Swati, you may culture in a liquid medium and incubate it in the shaker, or stir culture. Also, you can use a sonicator for a very short time (for liquid culture).
Good luck
Asking such a question, - first of all, - give the name of the fungus/wood pair!
In general, design of appropriate dry fermentation unit seems to be the easiest way.
For example; sterilize sawdust and hay separately in a baker at 105°C for 2 hours and let them cool in the desiccator. Be careful, do not shred the hay into small pieces! Majority of its fragments must be more than 1 cm. Then wet the sawdust with appropriate amounts of sterile distilled water [for example 20 to 1 w/v] and mix up with dry hay ( for example 3:1 w/w) before filling the fermentor.
Inoculation should be made appropriately to the properties of the fungus. Most probably, a small fragment of mycelium suits. Wait until the fungus colonizes the whole material, then let dry the material in the first step at open air 1-2 days, and subsequently finish drying either in desiccator over calcium chloride or choose a lyophilizer.
The dried material you can micronize easily. There is a legion of possibilities.
The powderized preparate can be easily suspended in water containing appropriate detergent (for example 1:1 Tween 20 and Tween 80 200 mg/L).
The standardization might be done by counting the living propagules per g of dry preparate, which process can be carried out by traditional two fold dilution technique. Use three series!
Unfortunately, these fungal species have good survival capacity.
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